Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Duplex dPCR System for Rapid Identification of Gram-Negative Pathogens in the Blood of Patients with Bloodstream Infection: A Culture-Independent Approach

  • Shin, Juyoun (Department of Microbiology, The Catholic University of Korea, College of Medicine) ;
  • Shin, Sun (Department of Microbiology, The Catholic University of Korea, College of Medicine) ;
  • Jung, Seung-Hyun (Department of Biochemistry, The Catholic University of Korea, College of Medicine) ;
  • Park, Chulmin (Vaccine Bio Research Institute, The Catholic University of Korea, College of Medicine, Seoul St. Mary's Hospital) ;
  • Cho, Sung-Yeon (Vaccine Bio Research Institute, The Catholic University of Korea, College of Medicine, Seoul St. Mary's Hospital) ;
  • Lee, Dong-Gun (Vaccine Bio Research Institute, The Catholic University of Korea, College of Medicine, Seoul St. Mary's Hospital) ;
  • Chung, Yeun-Jun (Department of Microbiology, The Catholic University of Korea, College of Medicine)
  • Received : 2021.03.26
  • Accepted : 2021.09.09
  • Published : 2021.11.28
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Abstract

Early and accurate detection of pathogens is important to improve clinical outcomes of bloodstream infections (BSI), especially in the case of drug-resistant pathogens. In this study, we aimed to develop a culture-independent digital PCR (dPCR) system for multiplex detection of major sepsis-causing gram-negative pathogens and antimicrobial resistance genes using plasma DNA from BSI patients. Our duplex dPCR system successfully detected nine targets (five bacteria-specific targets and four antimicrobial resistance genes) through five reactions within 3 hours. The minimum detection limit was 50 ag of bacterial DNA, suggesting that 1 CFU/ml of bacteria in the blood can be detected. To validate the clinical applicability, cell-free DNA samples from febrile patients were tested with our system and confirmed high consistency with conventional blood culture. This system can support early identification of some drug-resistant gram-negative pathogens, which can help improving treatment outcomes of BSI.

Keywords

Acknowledgement

This study was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by Ministry of Health & Welfare, Republic of Korea (Grant number: H14C2658 and HI14C3417) and National Research Foundation of Korea (2015M3C7A1064778, 2019R1A5A2027588, 2017M3C9A6047615).

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